Approaches to the Study of Somatostatin Biosynthesis
Part of the
Advances in Experimental Medicine and Biology
book series (AEMB, volume 188)
Our current knowledge of the processes regulating somatostatin biosynthesis is still scarce. Approaches used for the direct investigation of somatostatin biosynthesis in different tissues include 1) analysis of incorporation of labeled amino acids into somatostatin-like immunoreactivity (SLI), 2) cell-free translation of mRNA isolated from SLI producing tissues and 3) analysis of mRNA coding for somatostatin by cDNA blot hybridization.
Amino acid incorporation into SLI has been studied in a variety of systems, including anglerfish and rat pancreas, frog retina, rat dorsal root ganglia, cerebral cortex and hypothalamus. We have studied the neuronal biosynthesis of somatostatin using monolayer cultures of neonatal rat hypothalamic cells. Following pulse labeling with [3H]phenylalanine, the cellular extracts contained material that bound specifically to an immobilized anti-somatostatin antibody. Analysis of the bound label by gel chromatography and HPLC provided evidence for the presence of labeled somatostatin-14 (S-14), somatostatin-28 (S-28) and a precursor molecule of Mr 15,000 (15 K SLI). Pulse-chase experiments demonstrated a transfer of label from 15K SLI to material corresponding to S-28 and S-14.
Using cloned cDNAs complementary to somatostatin mRNA, the presence of somatostatin mRNA has been demonstrated in anglerfish pancreas and intestine, rat hypothalamus and antrum, as well as in a rat medullary thyroid carcinoma and a rat pancreatic cell line. We have recently studied the developmental regulation of somatostatin gene expression in the rat brain and stomach. Messenger RNA hybridizing specifically to a rat somatostatin cDNA probe was already clearly detectable in tissue extracts derived from brains of one week old rat fetuses. A marked increase of somatostatin mRNA occured between day 14 and day 21 of embryonic life. By contrast, in tissue extracts derived from stomach, somatostatin mRNA remained undetectable until shortly before birth. These marked differences in the tissue specific regulation of somatostatin gene expression during ontogenesis may reflect basic differences in the developmental regulation of somatostatin gene expression in neural vs. nonneural tissues or may be related to the onset of functional activity in the organs studied.
KeywordsSodium Butyrate Label Amino Acid Label Incorporation Chase Period Cell Free Translation
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